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Biotechnology and Bioengineering

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https://www.readbyqxmd.com/read/28627778/engineering-cho-cells-with-an-oncogenic-kit-improves-cells-growth-resilience-to-stress-and-productivity
#1
Mohamed Mahameed, Boaz Tirosh
An optimized biomanufacturing process in mammalian cells is contingent on the ability of the producing cells to reach high viable cell densities. In addition, at the peak of growth, cells need to continue producing the biological entity at a consistent quality. Thus, engineering cells with robust growth performance and resilience to variable stress conditions is highly desirable. The tyrosine kinase receptor, KIT, plays a key role in cell differentiation and the survival of several immune cell types. Its oncogenic mutant, D816V, endows cells with high proliferation capacity and resistance to kinase inhibitors...
June 19, 2017: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/28627740/biofabrication-of-soft-tissue-templates-for-engineering-the-bone-ligament-interface
#2
Ella Harris, Yurong Liu, Grainne Cuniffe, David Morrissey, Simon Carroll, Kevin Mulhall, Daniel J Kelly
Regenerating damaged tissue interfaces remains a significant clinical challenge, requiring recapitulation of the structure, composition and function of the native enthesis. In the ligament-to-bone interface this region transitions from ligament to fibrocartilage, to calcified cartilage and then to bone. This gradation in tissue types facilitates the transfer of load between soft and hard structures while minimizing stress concentrations at the interface. Previous attempts to engineer the ligament-bone interface have utilized various scaffold materials with an array of various cell types and/or biological cues...
June 19, 2017: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/28627739/use-of-a-protein-engineering-strategy-to-overcome-limitations-in-the-production-of-difficult-to-express-recombinant-proteins
#3
Hirra Hussain, David I Fisher, W Mark Abbott, Robert G Roth, Alan J Dickson
Certain recombinant proteins are deemed "difficult to express" in mammalian expression systems requiring significant cell and/or process engineering to abrogate expression bottlenecks. With increasing demand for the production of recombinant proteins in mammalian cells, low protein yields can have significant consequences for industrial processes. To investigate the molecular mechanisms that restrict expression of recombinant proteins, naturally secreted model proteins were analysed from the tissue inhibitors of metalloproteinase (TIMP) protein family...
June 19, 2017: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/28627736/enzyme-free-cell-detachment-mediated-by-resonance-vibration-with-temperature-modulation
#4
Yuta Kurashina, Makoto Hirano, Chikahiro Imashiro, Kiichiro Totani, Jun Komotori, Kenjiro Takemura
Cell detachment is an essential process in adherent cell culture. However, trypsinization, which is the most popular detachment technique used in culture, damages cellular membranes. Reducing cellular membrane damage during detachment should improve the quality of cell culture. In this article, we propose an enzyme-free cell detachment method based on resonance vibration with temperature modulation. We developed a culture device that can excite a resonance vibration and control temperature. We then evaluated the cell detachment ratio and the growth response, observed the morphology and analyzed the cellular protein of the collected cells-mouse myoblast cell line (C2C12)...
June 19, 2017: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/28627734/role-of-cytoskeletal-mechanics-and-cell-membrane-fluidity-in-the-intracellular-delivery-of-molecules-mediated-by-laser-activated-carbon-nanoparticles
#5
Stefany Y Holguin, Caleb F Anderson, Naresh N Thadhani, Mark R Prausnitz
Exposure of cells and nanoparticles to near-infrared nanosecond pulsed laser light can lead to efficient intracellular delivery of molecules while maintaining high cell viability by a photoacoustic phenomenon known as transient nanoparticle energy transduction (TNET). Here, we examined the influence of cytoskeletal mechanics and plasma membrane fluidity on intracellular uptake of molecules and loss of cell viability due to TNET. We found that destabilization of actin filaments using latrunculin A led to greater uptake of molecules and less viability loss caused by TNET...
June 19, 2017: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/28627727/development-of-hgf-binding-aptamers-with-the-combination-of-g4-promoter-derived-aptamer-selection-and-in-silico-maturation
#6
Tomomi Yokoyama, Kaori Tsukakoshi, Wataru Yoshida, Taiki Saito, Kentaro Teramoto, Nasa Savory, Koichi Abe, Kazunori Ikebukuro
We describe the selection of aptamers based on bioinformatics-based approaches without Systematic Evolution of Ligands by EXponential enrichment (SELEX). SELEX is a potent method; however, it is time intensive and the PCR-amplification step, which is essential step for SELEX, leads to the loss of good aptamers. We have developed an aptamer-screening method, G4 promoter-derived aptamer selection (G4PAS), and an aptamer-improving method, in silico maturation (ISM). They are based on in silico sequence selection and computer assisted directed evolution, respectively...
June 19, 2017: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/28627725/proteomic-analysis-of-host-cell-protein-dynamics-in-the-supernatant-of-fc-fusion-protein-producing-cho-dg44-and-dukx-b11-cell-lines-in-batch-and-fed-batch-cultures
#7
Jin Hyoung Park, Jong Hwa Jin, In Jung Ji, Hyun Joo An, Jong Won Kim, Gyun Min Lee
Chinese hamster ovary (CHO) cells are the most widely used host cell lines for the commercial production of therapeutic proteins including Fc-fusion proteins. During the culture of recombinant CHO (rCHO) cells, host cell proteins (HCPs), secreted from viable cells and released from dead cells, accumulate extracellularly, potentially impairing product quality. In this study, the HCPs that accumulated extracellularly in batch and fed-batch cultures of Fc-fusion protein-producing rCHO cell lines (DG-Fc and DUKX-Fc) were identified and quantified using nanoflow liquid chromatography-tandem mass spectrometry (LC-MS/MS), followed by gene ontology and functional analysis...
June 19, 2017: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/28627713/process-development-of-human-multipotent-stromal-cell-microcarrier-culture-using-an-automated-high-throughput-microbioreactor
#8
Qasim A Rafiq, Mariana P Hanga, Thomas R J Heathman, Karen Coopman, Alvin W Nienow, David J Williams, Christopher J Hewitt
Microbioreactors play a critical role in process development as they reduce reagent requirements and can facilitate high-throughput screening of process parameters and culture conditions. Here we have demonstrated and explained in detail, for the first time, the amenability of the automated ambr15 cell culture microbioreactor system for the development of scalable adherent human mesenchymal multipotent stromal/stem cell (hMSC) microcarrier culture processes. This was achieved by first improving suspension and mixing of the microcarriers and then improving cell attachment thereby reducing the initial growth lag phase...
June 19, 2017: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/28627711/fermentation-strategy-for-second-generation-ethanol-production-from-sugarcane-bagasse-hydrolyzate-by-spathaspora-passalidarum-and-scheffersomyces-stipitis
#9
S C Nakanishi, L B Soares, L E Biazi, V M Nascimento, A C Costa, G J M Rocha, J L Ienczak
Alcoholic fermentation of released sugars in pretreatment and enzymatic hydrolysis of biomass is a central feature for second generation ethanol (E2G) production. Saccharomyces cerevisiae used industrially in the production of first generation ethanol (E1G) convert sucrose, fructose and glucose into ethanol. However, these yeasts have no ability to ferment pentose (xylose). Therefore, the present work has focused on E2G production by Scheffersomyces stipitis and Spathaspora passalidarum. The fermentation strategy with high pitch, cell recycle, fed-batch mode and temperature decrease for each batch were performed in a hydrolyzate obtained from a pretreatment at 130 °C with NaOH solution (1...
June 19, 2017: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/28627710/oil-and-eicosapentaenoic-acid-production-by-the-diatom-phaeodactylum-tricornutum-cultivated-outdoors-in-green-wall-panel-gwp%C3%A2-reactors
#10
Liliana Rodolfi, Natascia Biondi, Alessia Guccione, Niccolò Bassi, Massimo D'Ottavio, Gimena Arganaraz, Mario R Tredici
Phaeodactylum tricornutum is a widely studied diatom and has been proposed as a source of oil and polyunsaturated fatty acids (PUFA), particularly eicosapentaenoic acid (EPA). Recent studies indicate that lipid accumulation occurs under nutritional stress. Aim of this research was to determine how changes in nitrogen availability affect productivity, oil yield and fatty acid (FA) composition of P. tricornutum UTEX 640. After preliminary laboratory trials, outdoor experiments were carried out in 40-L GWP® reactors under different nitrogen regimes in batch...
June 19, 2017: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/28600898/investigation-of-acid-phosphatase-variants-for-the-synthesis-of-phosphate-monoesters
#11
Gábor Tasnádi, Michaela Zechner, Mélanie Hall, Kai Baldenius, Klaus Ditrich, Kurt Faber
The major drawback of using phosphatases for transphosphorylation reactions lies in product depletion caused by the natural hydrolytic activity of the enzymes. Variants of PhoC-Mm from Morganella morganii and NSAP-Eb from Escherichia blattae were studied for their ability to maintain a high product level in the transphosphorylation of various primary alcohols. A single amino acid exchange delivered phosphatase variant PhoC-Mm G92D, which was able to catalyze the phosphorylation of primary alcohols without any major hydrolysis of the formed phosphate esters...
June 10, 2017: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/28600895/metabolic-modeling-of-energy-balances-in-mycoplasma-hyopneumoniae-shows-that-pyruvate-addition-increases-growth-rate
#12
Tjerko Kamminga, Simen-Jan Slagman, Jetta J E Bijlsma, Vitor A P Martins Dos Santos, Maria Suarez-Diez, Peter J Schaap
Mycoplasma hyopneumoniae is cultured on large-scale to produce antigen for inactivated whole-cell vaccines against respiratory disease in pigs. However, the fastidious nutrient requirements of this minimal bacterium and the low growth rate make it challenging to reach sufficient biomass yield for antigen production. In this study, we sequenced the genome of M. hyopneumoniae strain 11 and constructed a high quality constraint-based genome-scale metabolic model of 284 chemical reactions and 298 metabolites. We validated the model with time-series data of duplicate fermentation cultures to aim for an integrated model describing the dynamic profiles measured in fermentations...
June 10, 2017: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/28600892/a-novel-free-ammonia-based-pretreatment-technology-to-enhance-anaerobic-methane-production-from-primary-sludge
#13
Wei Wei, Xu Zhou, Guo-Jun Xie, Haoran Duan, Qilin Wang
This study proposed a novel free ammonia (FA i.e. NH3 ) pretreatment technology to enhance anaerobic methane production from primary sludge for the first time. The solubilisation of primary sludge was substantially enhanced following 24 h FA pretreatment (250∼680 mg NH3 -N/L), by which the release of soluble chemical oxygen demand (SCOD) (i.e. 0.4 mg SCOD/mg VS added; VS: volatile solids) was approximately 10 times as much as that without pretreatment (i.e. 0.03 mg SCOD/mg VS added). Then, biochemical methane potential (BMP) tests demonstrated that FA pretreatment of 250∼680 mg NH3 -N/L was capable of enhancing anaerobic methane production while the digestion time was more than 7 days...
June 10, 2017: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/28600876/rerouting-of-carbon-flux-in-a-glycogen-mutant-of-cyanobacteria-assessed-via-isotopically-non-stationary-13-c-metabolic-flux-analysis
#14
John I Hendry, Charulata Prasannan, Fangfang Ma, K Benedikt Möllers, Damini Jaiswal, Madhuri Digmurti, Doug K Allen, Niels-Ulrik Frigaard, Santanu Dasgupta, Pramod P Wangikar
Cyanobacteria, which constitute a quantitatively dominant phylum, have attracted attention in biofuel applications due to favorable physiological characteristics, high photosynthetic efficiency and amenability to genetic manipulations. However, quantitative aspects of cyanobacterial metabolism have received limited attention. In the present study, we have performed isotopically non-stationary (13) C metabolic flux analysis (INST-(13) C-MFA) to analyze rerouting of carbon in a glycogen synthase deficient mutant strain (glgA-I glgA-II) of the model cyanobacterium Synechococcus sp...
June 10, 2017: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/28600873/bone-tissue-bioprinting-for-craniofacial-reconstruction
#15
REVIEW
Pallab Datta, Veli Ozbolat, Bugra Ayan, Aman Dhawan, Ibrahim T Ozbolat
Craniofacial (CF) tissue is an architecturally complex tissue consisting of both bone and soft tissues with significant patient specific variations. Conditions of congenital abnormalities, tumor resection surgeries, and traumatic injuries of the CF skeleton can result in major deficits of bone tissue. Despite advances in surgical reconstruction techniques, management of CF osseous deficits remains a challenge. Due its inherent versatility, bioprinting offers a promising solution to address these issues. In this review, we present and analyze the current state of bioprinting of bone tissue and highlight how these techniques may be adapted to serve regenerative therapies for CF applications...
June 10, 2017: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/28600864/co-utilization-of-hexoses-by-a-microconsortium-of-sugar-specific-e-coli-strains
#16
Todd C Chappell, Nikhil U Nair
Escherichia coli is an important commercial species used for production of biofuels, biopolymers, organic acids, sugar alcohols, and natural compounds. Processed biomass and agroindustrial byproducts serve as low-cost nutrient sources and contain a variety of hexoses available for bioconversion. However, metabolism of hexose mixtures by E. coli is inefficient due to carbon catabolite repression (CCR), where the transport and catabolic activity of one or more carbon sources is repressed and/or inhibited by the transport and catabolism of another carbon source...
June 10, 2017: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/28574594/a-novel-protein-engineered-hepatocyte-growth-factor-analog-released-via-a-shear-thinning-injectable-hydrogel-enhances-post-infarction-ventricular-function
#17
Amanda N Steele, Lei Cai, Vi N Truong, Bryan B Edwards, Andrew B Goldstone, Anahita Eskandari, Aaron C Mitchell, Laura M Marquardt, Abbygail A Foster, Jennifer R Cochran, Sarah C Heilshorn, Y Joseph Woo
In the last decade, numerous growth factors and biomaterials have been explored for the treatment of myocardial infarction (MI). While pre-clinical studies have demonstrated promising results, clinical trials have been disappointing and inconsistent, likely due to poor translatability. In the present study, we investigate a potential myocardial regenerative therapy consisting of a protein-engineered dimeric fragment of hepatocyte growth factor (HGFdf) encapsulated in a shear-thinning, self-healing, bioengineered hydrogel (SHIELD)...
June 2, 2017: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/28574582/production-of-g-protein-coupled-receptors-in-an-insect-based-cell-free-system
#18
Andrei Sonnabend, Viola Spahn, Marlitt Stech, Anne Zemella, Christoph Stein, Stefan Kubick
The biochemical analysis of human cell membrane proteins remains a challenging task due to the difficulties in producing sufficient quantities of functional protein. G protein-coupled receptors (GPCRs) represent a main class of membrane proteins and drug targets, which are responsible for a huge number of signaling processes regulating various physiological functions in living cells. To circumvent the current bottlenecks in GPCR studies, we propose the synthesis of GPCRs in eukaryotic cell-free systems based on extracts generated from insect (Sf21) cells...
June 2, 2017: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/28543036/self-assembly-of-two-hydrophobins-from-marine-fungi-affected-by-interaction-with-surfaces
#19
Paola Cicatiello, Principia Dardano, Marinella Pirozzi, Alfredo Maria Gravagnuolo, Luca De Stefano, Paola Giardina
Hydrophobins are amphiphilic fungal proteins endowed with peculiar characteristics, such as a high surface activity and an interface triggered self-assembly. Several applications of these proteins have been proposed in the food, cosmetics and biomedical fields. Moreover, their use as proteinaceous coatings can be effective for materials and nanomaterials applications. The discovery of novel hydrophobins with diverse properties may be advantageous from both the scientific and industrial points of view. Stressful environmental conditions of fungal growth may induce the production of proteins with peculiar features...
May 22, 2017: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/28542804/assessment-of-an-engineered-endothelium-via-single-photon-emission-computed-tomography
#20
Bin Jiang, Yidi Wu, Chad R Haney, Chongwen Duan, Guillermo A Ameer
The clinical translation of cell-based therapeutics often requires highly sensitive, non-invasive imaging tools to assess cell function and distribution in vivo. The objective of this research was to determine whether human Sodium-Iodide Symporter (hNIS) ectopic expression in endothelial cells (ECs) in combination with single-photon emission computed tomography (SPECT) is a feasible approach to non-invasively monitor the presence and viability of an engineered endothelium on expanded polytetrafluoroethylene (ePTFE)...
May 22, 2017: Biotechnology and Bioengineering
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