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Biotechnology Journal

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https://www.readbyqxmd.com/read/28514082/universal-label-free-in-process-quantification-of-influenza-virus-like-particles
#1
Sofia B Carvalho, Mafalda G Moleirinho, David Wheatley, John Welsh, René Gantier, Paula M Alves, Cristina Peixoto, Manuel J T Carrondo
Virus-like particles (VLPs) are becoming established as vaccines, in particular for influenza pandemics, increasing the interest in the development of VLPs manufacturing bioprocess. However, for complex VLPs, the analytical tools used for quantification are not yet able to keep up with the bioprocess progress. Currently, quantification for Influenza relies on traditional methods: hemagglutination assay or Single Radial Immunodiffusion. These analytical technologies are time-consuming, cumbersome, and not supportive of efficient downstream process development and monitoring...
May 17, 2017: Biotechnology Journal
https://www.readbyqxmd.com/read/28504349/polysome-profiling-of-mab-producing-cho-cell-lines-links-translational-control-of-cell-proliferation-and-recombinant-mrna-loading-onto-ribosomes-with-global-and-recombinant-protein-synthesis
#2
Charlotte L Godfrey, Emma J Mead, Olalekan Daramola, Sarah Dunn, Diane Hatton, Ray Field, Gary Pettman, C Mark Smales
mRNA translation is a key process determining growth, proliferation and duration of a Chinese hamster ovary (CHO) cell culture and influences recombinant protein synthesis rate. During bioprocessing, CHO cells can experience stresses leading to reprogramming of translation and decreased global protein synthesis. Here we apply polysome profiling to determine reprogramming and translational capabilities in host and recombinant monoclonal antibody-producing (mAb) CHO cell lines during batch culture. Recombinant cell lines with the fastest cell specific growth rates were those with the highest global translational efficiency...
May 15, 2017: Biotechnology Journal
https://www.readbyqxmd.com/read/28497573/simplex-based-optimization-of-numerical-and-categorical-inputs-in-early-bioprocess-development-case-studies-in-ht-chromatography
#3
Spyridon Konstantinidis, Nigel Titchener-Hooker, Ajoy Velayudhan
Bioprocess development studies often involve the investigation of numerical and categorical inputs via the adoption of Design of Experiments (DoE) techniques. An attractive alternative is the deployment of a grid compatible Simplex variant which has been shown to yield optima rapidly and consistently. In this work, the method is combined with dummy variables and it is deployed in three case studies wherein spaces are comprised of both categorical and numerical inputs, a situation intractable by traditional Simplex methods...
May 12, 2017: Biotechnology Journal
https://www.readbyqxmd.com/read/28497498/cryoprotection-and-banking-of-living-cells-in-a-3d-multiple-emulsion-based-carrier
#4
Ewa Dluska, Zhanfeng Cui, Agnieszka Markowska-Radomska, Agata Metera, Konrad Kosicki
This paper addresses the problem of the low viability of cells during traditional cryopreservation by developing multiple emulsion-based carriers for the encapsulation and cryopreservation of cells. Multiple emulsions are hierarchically structured dispersed systems of "droplets in drops", which allow bioactive ingredients to be encapsulated. The problem of low survival rates refers to the stage of their freezing-thawing in traditional suspensions. During freezing-thawing processes, irreversible damage to cells occurs due to the formation of ice crystals, cell dehydration, and cryoprotectants' toxicity...
May 12, 2017: Biotechnology Journal
https://www.readbyqxmd.com/read/28488793/atomic-force-microscopy-studies-of-bioprocess-engineering-surfaces-imaging-interactions-and-mechanical-properties-mediating-bacterial-adhesion
#5
REVIEW
Sean A James, Nidal Hilal, Chris J Wright
The detrimental effect of bacterial biofilms on process engineering surfaces is well documented. Thus, interest in the early stages of bacterial biofilm formation; in particular bacterial adhesion and the production of anti-fouling coatings has grown exponentially as a field. During this time, Atomic force microscopy (AFM) has emerged as a critical tool for the evaluation of bacterial adhesion. Due to its versatility AFM offers not only insight into the topographical landscape and mechanical properties of the engineering surfaces, but elucidates, through direct quantification the topographical and biomechnical properties of the foulants The aim of this review is to collate the current research on bacterial adhesion, both theoretical and practical, and outline how AFM as a technique is uniquely equipped to provide further insight into the nanoscale world at the bioprocess engineering surface...
May 10, 2017: Biotechnology Journal
https://www.readbyqxmd.com/read/28488758/engineered-humicola-insolens-cutinase-for-efficient-cellulose-acetate-deacetylation
#6
Abhijit N Shirke, Glenn L Butterfoss, Rakhi Saikia, Aditya Basu, Leonardo de Maria, Allan Svendsen, Richard A Gross
Cutinases comprise a family of esterases with broad hydrolytic activity for chain and pendant ester groups. This work aimed to identify and improve an efficient cutinase for cellulose acetate (CA) deacetylation. The development of a mild method for CA fiber surface deacetylation will result in improved surface hydrophilicity and reactivity while, when combined with cellulases, a route to the full recycling of CA to acetate and glucose. In this study, the comparative CA deacetylation activity of 4 homologous wild-type (wt) fungal cutinases from Aspergillus oryzae (AoC), Thiellavia terrestris (TtC), Fusarium solani (FsC), and Humicola insolens (HiC) was determined by analysis of CA deacetylation kinetics...
May 10, 2017: Biotechnology Journal
https://www.readbyqxmd.com/read/28481000/conscious-coupling-the-challenges-and-opportunities-of-cascading-enzymatic-microreactors
#7
REVIEW
Pia Gruber, Marco P C Marques, Brian O'Sullivan, Frank Baganz, Roland Wohlgemuth, Nicolas Szita
The continuous production of high value or difficult to synthesize products is of increasing interest to the pharmaceutical industry. Cascading reaction systems have already been employed for chemical synthesis with great success, allowing a quick change in reaction conditions and addition of new reactants as well as removal of side products. A cascading system can remove the need for isolating unstable intermediates, increasing the yield of a synthetic pathway. Based on the success for chemical synthesis, the question arises how cascading systems could be beneficial to chemo-enzymatic or biocatalytic synthesis...
May 8, 2017: Biotechnology Journal
https://www.readbyqxmd.com/read/28464535/compartmentalized-metabolic-engineering-for-biochemical-and-biofuel-production
#8
REVIEW
Herbert M Huttanus, Xueyang Feng
Sub-cellular compartments create specialized reaction chambers in eukaryotes. These compartments provide favorable micro-environments for many metabolic processes. Recently, metabolic engineers have explored the concept of pathway compartmentalization to enhance the performance of metabolic pathways. This strategy offers many unique advantages, including (i) increased local concentrations of enzymes and substrates, (ii) accessing alternate substrate pools, (iii) separation from competing reactions, and (iv) isolation of harmful intermediates or conditions needed for the pathway...
May 2, 2017: Biotechnology Journal
https://www.readbyqxmd.com/read/28449226/an-integrated-network-analysis-identifies-how-arcab-enables-metabolic-oscillations-in-the-nitric-oxide-detoxification-network-of-escherichia-coli
#9
Sarah A Sacco, Kristin J Adolfsen, Mark P Brynildsen
The virulences of many pathogens depend on their abilities to detoxify the immune antimicrobial nitric oxide (NO•). The functions of bacterial NO• detoxification machinery depend on oxygen (O2 ), with O2 inhibiting some enzymes, whereas others use it as a substrate. Previously, Escherichia coli NO• detoxification was found to be highly attenuated under microaerobic conditions and metabolic oscillations were observed. The oscillations in [NO•] and [O2 ] were found to result from the inhibitory action of NO• on aerobic respiration, the catalytic inactivation of NO• by Hmp (an NO• dioxygenase), and an imbalanced competition for O2 between Hmp and cytochrome terminal oxidase activity...
April 27, 2017: Biotechnology Journal
https://www.readbyqxmd.com/read/28439989/using-size-controlled-multicellular-spheroids-of-murine-adenocarcinoma-cells-to-efficiently-establish-pulmonary-tumors-in-mice
#10
Tomoko Nishikawa, Yutaro Tanaka, Makiya Nishikawa, Kosuke Kusamori, Narumi Mizuno, Yuya Mizukami, Yuka Ogino, Kazunori Shimizu, Satoshi Konishi, Yuki Takahashi, Yoshinobu Takakura
Previous studies demonstrated that multicellular spheroids developed using polydimethylsiloxane-based microwells exhibited superior functions, such as insulin secretion from pancreatic cells, over suspended cells. To successfully apply these spheroids, the effect of spheroid size on cellular functions must be determined. In this study, using murine adenocarcinoma colon26 cells, we examined whether such spheroids were useful for developing tumor-bearing animal models, which requires the efficient and stable engraftment of cancer cells at implanted sites and/or metastatic sites...
April 25, 2017: Biotechnology Journal
https://www.readbyqxmd.com/read/28430399/targeting-conformational-activation-of-cdk2-kinase
#11
Morgan Pellerano, Sergey Tcherniuk, Corine Perals, Thi Nhu Ngoc Van, Elsa Garcin, Florence Mahuteau-Betzer, Marie-Paule Teulade-Fichou, May C Morris
Cyclin-dependent kinases constitute attractive pharmacological targets for cancer therapeutics, yet inhibitors in clinical trials target the ATP-binding pocket of the CDK and therefore suffer from limited selectivity and emergence of resistance. The more recent development of allosteric inhibitors targeting conformational plasticity of protein kinases offers promising perspectives for therapeutics. In particular tampering with T-loop dynamics of CDK2 kinase would provide a selective means of inhibiting this kinase, by preventing its conformational activation...
April 21, 2017: Biotechnology Journal
https://www.readbyqxmd.com/read/28429856/glycosyltransferase-cascades-for-natural-product-glycosylation-use-of-plant-instead-of-bacterial-sucrose-synthases-improves-the-udp-glucose-recycling-from-sucrose-and-udp
#12
Alexander Gutmann, Alexander Lepak, Margo Diricks, Tom Desmet, Bernd Nidetzky
Natural product glycosylations by Leloir glycosyltransferases (GTs) require expensive nucleotide-activated sugars as substrates. Sucrose synthase (SuSy) converts sucrose and uridine 5'-diphosphate (UDP) into UDP-glucose. Coupling of SuSy and GT reactions in one-pot cascade transformations creates a UDP cycle, which regenerates the UDP-glucose continuously and so makes it an expedient donor for glucoside production. Here we compare SuSys with divergent kinetic characteristics for UDP-glucose recycling in the synthesis of the natural C-glucoside nothofagin...
April 21, 2017: Biotechnology Journal
https://www.readbyqxmd.com/read/28429845/following-nature-s-roadmap-folding-factors-from-plasma-cells-led-to-improvements-in-antibody-secretion-in-s-cerevisiae
#13
Essi V Koskela, Jorg C de Ruijter, Alexander D Frey
Therapeutic protein production in yeast is a reality in industry with an untapped potential to expand to more complex proteins, such as full-length antibodies. Despite multiple numerous engineering approaches, cellular limitations are preventing the use of Saccharomyces cerevisiae as the titers of recombinant antibodies are currently not competitive. Instead of a host specific approach, we demonstrate the possibility of adopting the features from native producers of antibodies, plasma cells, to improve antibody production in yeast...
April 21, 2017: Biotechnology Journal
https://www.readbyqxmd.com/read/28418108/metabolic-characterization-and-transformation-of-the-non-dairy-lactococcus-lactis-strain-kf147-for-production-of-ethanol-from-xylose
#14
Kia Vest Petersen, Jianming Liu, Jun Chen, Jan Martinussen, Peter Ruhdal Jensen, Christian Solem
The non-dairy lactic acid bacterium Lactococcus lactis KF147 can utilize xylose as the sole energy source. To assess whether KF147 could serve as a platform organism for converting second generation sugars into useful chemicals, we characterized growth and product formation for KF147 when grown on xylose. In a defined medium KF147 was found to co-metabolize xylose and arginine, resulting in bi-phasic growth. Especially at low xylose concentrations, arginine significantly improved growth rate. To facilitate further studies of the xylose metabolism, we eliminated arginine catabolism by deleting the arcA gene encoding the arginine deiminase...
April 18, 2017: Biotechnology Journal
https://www.readbyqxmd.com/read/28403549/chaotropic-heat-treatment-resolves-native-like-aggregation-of-a-heterologously-produced-hyperthermostable-laminarinase
#15
Adrie H Westphal, Astrid A Geerke-Volmer, Carlo P M van Mierlo, Willem J H van Berkel
Production of hyperthermostable enzymes in mesophilic hosts frequently causes undesired aggregation of these proteins. During production of Pyrococcus furiosus endo-β-1,3 glucanase (LamA) in Escherichia coli, soluble and insoluble species form. Here, we address the composition of this mixture, including the nature of LamA conformers, and establish a method to increase the yield of native monomer. With gel electrophoresis, size-exclusion chromatography, light scattering, circular dichroism and enzyme kinetics we show that approximately 50 % of heterologously produced LamA is soluble, and that 40 % of this fraction constitutes native-like oligomers and non-native monomers...
April 12, 2017: Biotechnology Journal
https://www.readbyqxmd.com/read/28398709/continuous-desalting-of-refolded-protein-solution-improves-capturing-in-ion-exchange-chromatography-a-seamless-process
#16
Nicole Walch, Alois Jungbauer
Truly continuous biomanufacturing processes enable an uninterrupted feed stream throughout the whole production without the need for holding tanks. We have utilized microporous anion and cation exchangers into which only salts, but not proteins, can penetrate into the pores for desalting of protein solutions, while diafiltration or dilution is usually employed for feed adjustments. Anion exchange and cation exchange chromatography columns were connected in series to remove both anions and cations. To increase operation performance, a continuous process was developed comprised of four columns...
April 11, 2017: Biotechnology Journal
https://www.readbyqxmd.com/read/28397983/high-yield-production-of-recombinant-crm197-a-non-toxic-mutant-of-diphtheria-toxin-in-the-periplasm-of-escherichia-coli
#17
Philippe Goffin, Marianne Dewerchin, Philippe De Rop, Normand Blais, Philippe Dehottay
A high cell density fed-batch process was developed for production of recombinant CRM197, a non-toxic mutant of diphtheria toxin widely used as a carrier in polysaccharide-protein conjugate vaccines. Fully soluble recombinant CRM197 was obtained in high yields and with an authentic N-terminus, by targeting the protein to the periplasm of Escherichia coli using the Signal Recognition Particle (SRP)-dependent signal sequence of FlgI. Response Surface Methodology (RSM) was used to optimize the set-points of key process parameters (pH and feed rate at induction)...
April 11, 2017: Biotechnology Journal
https://www.readbyqxmd.com/read/28371407/biocatalytic-virus-capsid-as-nanovehicle-for-enzymatic-activation-of-tamoxifen-in-tumor-cells
#18
Alejandro Tapia-Moreno, Karla Juarez-Moreno, Oscar Gonzalez-Davis, Ruben D Cadena-Nava, Rafael Vazquez-Duhalt
Most of the drugs used in chemotherapy should be activated by a transformation catalyzed by cytochrome P450 (CYP) enzymes. In this work, bacteriophage P22 virus-like particles (VLPs) containing CYP activity, immunologically inert and functionalized in order to be recognized by human cervix carcinoma cells and human breast adenocarcinoma cells were designed. The CYP was encapsulated inside the virus capsid obtained from the bacteriophage P22. CYP and coat protein were both heterologously expressed in E. coli...
April 3, 2017: Biotechnology Journal
https://www.readbyqxmd.com/read/28371347/genetic-engineering-of-the-branched-fatty-acid-metabolic-pathway-of-bacillus-subtilis-for-the-overproduction-of-surfactin-c14-isoform
#19
Debarun Dhali, François Coutte, Anthony Argüelles Arias, Sandrine Auger, Vladimir Bidnenko, Gabrielle Chataigné, Michael Lalk, Joachim Niehren, Joana de Sousa, Cristian Versari, Philippe Jacques
Surfactin, a lipopeptide produced by Bacillus subtilis, is one of the most powerful biosurfactants known. This molecule consists of a cyclic heptapeptide linked to a β-hydroxy fatty acid chain. The isomery and the length of the fatty acid (FA) chain are responsible for the surfactin's activities. In this study, the gene codY, which encode for the global transcriptional regulator and the gene lpdV, located in the bkd operon (lpdV, bkdAA, bkdAB and bkdB genes), which is responsible for the last step of the branched chain amino acid (BCAA) degradation in acyl-CoA were deleted...
April 3, 2017: Biotechnology Journal
https://www.readbyqxmd.com/read/28371300/a-single-step-facs-sorting-strategy-in-conjunction-with-fluorescent-vital-dye-imaging-efficiently-assures-clonality-of-biopharmaceutical-production-cell-lines
#20
Jürgen Fieder, Patrick Schulz, Ingo Gorr, Harald Bradl, Till Wenger
The development of biopharmaceutical production cell lines typically starts with generation of heterogeneous populations of cells, from which then single cell clones are established. Several regulatory guidelines require that production cell lines are clonal, and the actual demonstration of clonality has been increasingly demanded by regulatory authorities over the last years. Here, the authors describe the relative contribution of flow cytometry mediated deposition of single cells in multiwell plates and subsequent imaging to assurance of clonality in a state of the art approach to single cell generation...
April 3, 2017: Biotechnology Journal
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