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Biotechnology Journal

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https://www.readbyqxmd.com/read/29345424/a-fluorescence-based-sensor-assay-that-monitors-general-protein-aggregation-in-human-cells
#1
Marisa Pereira, Diogo Tomé, Ana S Domingues, Ana S Varanda, Cristiana Paulo, Manuel A S Santos, Ana R Soares
Protein conformational disorders are characterized by disruption of protein folding and toxic accumulation of protein aggregates. Here we describe a sensitive and simple method to follow and monitor general protein aggregation in human cells. Heat shock protein 27 (HSP27) is an oligomeric small heat shock protein that binds and keeps unfolded proteins in a folding competent state. This high specificity of HSP27 for aggregated proteins can be explored to monitor aggregation in living cells by fusing it to a fluorescent protein as Green Fluorescent Protein (GFP)...
January 18, 2018: Biotechnology Journal
https://www.readbyqxmd.com/read/29345420/integration-of-gas-enhanced-oil-recovery-in-multiphase-fermentations-for-the-microbial-production-of-fuels-and-chemicals
#2
Susana Pedraza-de la Cuesta, Lore Keijzers, Luuk A M van der Wielen, Maria C Cuellar
In multiphase fermentations where the product forms a second liquid phase or where solvents are added for product extraction, turbulent conditions disperse the oil phase as droplets. Surface-active components (SACs) present in the fermentation broth can stabilize the product droplets thus forming an emulsion. Breaking this emulsion increases process complexity and consequently the production cost. In previous works, it has been proposed to promote demulsification of oil/supernatant emulsions in an off-line batch bubble column operating at low gas flow rate...
January 18, 2018: Biotechnology Journal
https://www.readbyqxmd.com/read/29341493/modeling-the-downstream-processing-of-monoclonal-antibodies-reveals-cost-advantages-for-continuous-methods-for-a-broad-range-of-manufacturing-scales
#3
Jonathan Hummel, Mark Pagkaliwangan, Xhorxhi Gjoka, Terence Davidovits, Rick Stock, Thomas Ransohoff, Rene Gantier, Mark Schofield
The biopharmaceutical industry is evolving in response to changing market conditions, including increasing competition and growing pressures to reduce costs. Single-use technologies and continuous bioprocessing have attracted attention as potential facilitators of cost-optimized manufacturing for monoclonal antibodies. While disposable bioprocessing has been adopted at many scales of manufacturing, continuous bioprocessing has yet to reach the same level of implementation. In this study, the cost of goods of Pall Life Science's integrated, continuous bioprocessing platform was modeled, along with that of purification processes in stainless-steel and single-use batch formats...
January 17, 2018: Biotechnology Journal
https://www.readbyqxmd.com/read/29341427/lc-ms-based-intact-igg-and-released-glycan-analysis-for-bioprocessing-applications
#4
Tianhua Wang, Kong Meng Hoi, Henning Stöckmann, Corrine Wan, Lyn Chiin Sim, Noor Hayati Bte Kamari, Shi Jie Tay, Ce Huang Poo, Susanto Woen, Yuangsheng Yang, Peiqing Zhang, Pauline M Rudd
Robust plate based antibody glycan analysis platforms are urgently needed for biopharmaceutical development and manufacturing as well as for clinical biomarker research. A 96-well plate based workflow has been developed to analyze both intact IgG antibodies and released N-glycans using an Orbitrap Fusion Mass Spectrometer and an LC/MS method on the Waters UNIFI platform. Here we describe such a workflow including protein A purification, PNGaseF digestion, 2-AB labeling and SPE clean-up. The measured IgG glycan profile is consistent with that obtained from non-plate based method and commercial kit and has the advantage of less hands-on time...
January 17, 2018: Biotechnology Journal
https://www.readbyqxmd.com/read/29337429/co-expression-of-orfcma-with-phb-depolymerase-phazcma-in-escherichia-coli-induces-efficient-whole-cell-biodegradation-of-polyesters
#5
Ming-Chieh Lee, En-Jung Liu, Cheng-Han Yang, Li-Jung Hsiao, Tzong-Ming Wu, Si-Yu Li
Whole-cell degradation of polyesters not only avoids the tedious process of enzyme separation, but also allows the degraded product to be reused as a carbon source. In this study, Escherichia coli BL21(DE3) harboring phaZCma , a gene encoding poly(3-hydroxybutyrate) (PHB) depolymerase from Caldimonas manganoxidans, was constructed. The extra-cellular fraction of E. coli/pPHAZ exhibited a fast PHB degradation rate where it only took 35 h to completely degrade PHB films, while C. manganoxidans took 81 h to do the same...
January 16, 2018: Biotechnology Journal
https://www.readbyqxmd.com/read/29334181/donor-variability-in-growth-kinetics-of-healthy-hmscs-using-manual-processing-considerations-for-manufacture-of-cell-therapies
#6
Giulia Detela, Owen W Bain, Hae-Won Kim, David J Williams, Chris Mason, Anthony Mathur, Ivan B Wall
Human mesenchymal stromal cells (hMSCs) are excellent candidates for cell therapy but their expansion to desired clinical quantities can be compromised by ex vivo processing, due to differences between donor material and process variation. The aim of this article was to characterize growth kinetics of healthy baseline "reference" hMSCs using typical manual processing. Bone-marrow derived hMSCs from ten donors were isolated based on plastic adherence, expanded and analyzed for their growth kinetics until passage 4...
January 15, 2018: Biotechnology Journal
https://www.readbyqxmd.com/read/29330927/defined-serum-free-medium-for-bioreactor-culture-of-an-immortalized-human-erythroblast-cell-line
#7
Esmond Lee, Zhong Ri Lim, Hong Yu-Chen, Bin Xia Yang, Alan Tin-Lun Lam, Allen Kuan-Liang Chen, Jaichandran Sivalingam, Shaul Reuveny, Yuin-Han Loh, Steve Kah-Weng Oh
Anticipated shortages in donated blood supply have prompted investigation of alternative approaches for in-vitro production of red blood cells (RBCs), such as expansion of conditional immortalization erythroid progenitors. However, there is a bioprocessing challenge wherein factors promoting maximal cell expansion and growth-limiting inhibitory factors are yet to be investigated. We used an erythroblast cell line (ImEry) derived from immortalizing CD71+CD235a+ erythroblast from adult peripheral blood for optimization of expansion culture conditions...
January 13, 2018: Biotechnology Journal
https://www.readbyqxmd.com/read/29328552/changes-in-chromosome-counts-and-patterns-in-cho-cell-lines-upon-generation-of-recombinant-cell-lines-and-subcloning
#8
Sabine Vcelar, Michael Melcher, Norbert Auer, Astrid Hrdina, Anja Puklowski, Friedrich Leisch, Vaibhav Jadhav, Till Wenger, Martina Baumann, Nicole Borth
Chinese hamster ovary cells are the number one production system for therapeutic proteins. A pre-requirement for their use in industrial production of biopharmaceuticals is to be clonal, thus originating from a single cell in order to be phenotypically and genomically identical. In the present study it was evaluated whether standard procedures, such as the generation of a recombinant cell line in combination with selection for a specific and stable phenotype (expression of the recombinant product) or subcloning have any impact on karyotype stability or homogeneity in CHO cells...
January 12, 2018: Biotechnology Journal
https://www.readbyqxmd.com/read/29328545/computer-simulation-elucidates-yeast-flocculation-and-sedimentation-for-efficient-industrial-fermentation
#9
Chen-Guang Liu, Zhi-Yang Li, Yue Hao, Juan Xia, Feng-Wu Bai, Muhammad Aamer Mehmood
Flocculation plays an important role in the immobilized fermentation of biofuels and biochemicals. It is essential to understand the flocculation phenomenon at physical and molecular scale; however, flocs can't be studied directly due to fragile nature. Hence, the present study was focused on the morphological specificities of yeast flocs formation and sedimentation via the computer simulation by a single floc growth model, based on Diffusion-Limited Aggregation (DLA) model. The impact of shear force, adsorption, and cell propagation on porosity and floc size was systematically illustrated...
January 12, 2018: Biotechnology Journal
https://www.readbyqxmd.com/read/29323472/integrated-analysis-of-the-transcriptome-and-metabolome-of-corynebacterium-glutamicum-during-penicillin-induced-glutamic-acid-production
#10
Takashi Hirasawa, Masaki Saito, Katsunori Yoshikawa, Chikara Furusawa, Hiroshi Shmizu
Corynebacterium glutamicum is known for its ability to produce glutamic acid and has been utilized for the fermentative production of various amino acids. Glutamic acid production in C. glutamicum is induced by penicillin. In this study, we analyzed the transcriptome and metabolome of C. glutamicum to understand the mechanism of penicillin-induced glutamic acid production. Transcriptomic analysis with DNA microarray revealed that expression of some glycolysis- and TCA cycle-related genes, which include those encoding the enzymes involved in conversion of glucose to 2-oxoglutaric acid, was upregulated after penicillin addition...
January 11, 2018: Biotechnology Journal
https://www.readbyqxmd.com/read/29323465/multiplexed-digital-mrna-expression-analysis-profiles-system-wide-changes-in-mrna-abundance-and-responsiveness-of-upr-specific-gene-expression-changes-during-batch-culture-of-recombinant-chinese-hamster-ovary-cells
#11
Rodrigo Maldonado-Agurto, Alan J Dickson
The unfolded protein response (UPR) signalling pathway is viewed as critical for setting the effectiveness of recombinant protein expression in CHO cells. In this study, we used Nanostring nCounter technology to study expression of a group of genes associated with cellular processes linked to UPR activation under ER stress and the changing environment of a batch culture. Time course induction of ER stress, using tunicamycin (TM), showed a group of genes such as Chop, Trb3, Sqstm1, Grp78 and Herpud1 responded rapidly to TM inhibition of N-glycosylation, whilst others such as Atf5, Odz4 and Birc5 exhibited a delayed response...
January 11, 2018: Biotechnology Journal
https://www.readbyqxmd.com/read/29319229/-cyt-nuc-a-customizable-and-documenting-imagej-macro-for-evaluation-of-protein-distributions-between-cytosol-and-nucleus
#12
Tilman Grune, Richard Kehm, Annika Höhn, Tobias Jung
Large amounts of data from multi-channel, high resolution, fluorescence microscopic images require tools that provide easy, customizable and reproducible high-throughput analysis. The freeware "ImageJ" has become one of the standard tools for scientific image analysis. Since ImageJ offers recording of "macros", even a complex multi-step process can be easily applied fully automated to large numbers of images, saving both time and reducing human subjective evaluation. In this work, we present "Cyt/Nuc", an ImageJ macro, able to recognize and to compare the nuclear and cytosolic areas of tissue samples, in order to investigate distributions of immunostained proteins between both compartments, while it documents in detail the whole process of evaluation and pattern recognition...
January 10, 2018: Biotechnology Journal
https://www.readbyqxmd.com/read/29316330/rna-seq-highlights-high-clonal-variation-in-monoclonal-antibody-producing-cho-cells
#13
Camila A Orellana, Esteban Marcellin, Robin W Palfreyman, Trent P Munro, Peter P Gray, Lars K Nielsen
The development of next-generation sequencing technologies has opened new opportunities to better characterize complex eukaryotic cells. Chinese hamster ovary (CHO) cells play a primary role in therapeutic protein production, with currently five of the top ten blockbuster drugs produced in CHO. However, engineering superior CHO cells with improved production features has had limited success to date and cell lines are still developed through the generation and screening of large strain pools. Here, we applied RNA sequencing to contrast a high and a low monoclonal antibody producing cell line...
January 5, 2018: Biotechnology Journal
https://www.readbyqxmd.com/read/29316325/glycoengineering-in-cho-cells-advances-in-systems-biology
#14
REVIEW
Vijay Tejwani, Mikael R Andersen, Jong Hyun Nam, Susan T Sharfstein
For several decades, glycoprotein biologics have been successfully produced from Chinese hamster ovary (CHO) cells. The therapeutic efficacy and potency of glycoprotein biologics are often dictated by their post translational modifications, particularly glycosylation, which unlike protein synthesis, is a non-templated process. Consequently, both native and recombinant glycoprotein production generate heterogeneous mixtures containing variable amounts of different glycoforms. Stability, potency, plasma half-life, and immunogenicity of the glycoprotein biologic are directly influenced by the glycoforms...
January 5, 2018: Biotechnology Journal
https://www.readbyqxmd.com/read/29316283/split-cas9-not-hairs-advancing-the-therapeutic-index-of-crispr-technology
#15
REVIEW
Carolin Schmelas, Dirk Grimm
The discovery that the bacterial CRISPR/Cas9 system can be translated into mammalian cells continues to have an unprecedented impact on the biomedical research community, as it largely facilitates efforts to experimentally interrogate or therapeutically modify the cellular genome. In particular, CRISPR promises the ability to correct disease-associated genetic defects, or to target and destroy invading foreign DNA, in a simple, efficient and selective manner directly in affected human cells or tissues. Here, we highlight a set of exciting new strategies that aim at further increasing the therapeutic index of CRISPR technologies, by reducing the size of Cas9 expression cassettes and thus enhancing their compatibility with viral gene delivery vectors...
January 5, 2018: Biotechnology Journal
https://www.readbyqxmd.com/read/29316243/development-of-an-alginate-array-platform-to-decouple-the-effect-of-multiparametric-perturbations-on-human-pluripotent-stem-cells-during-pancreatic-differentiation
#16
Thomas C Richardson, Shibin Mathew, Joseph E Candiello, Saik K Goh, Prashant N Kumta, Ipsita Banerjee
Human embryonic stem cells (hESC)-derived functional cells hold great promise for regenerative cell therapy. Currently approved strategies for clinical translation requires the isolation of the hESCs-derived cells in materials allowing transfer of reagents but preventing integration with the host. However, hESC fate is known to be sensitive to its local microenvironment, both chemical and physical. Given the complexity of hESC response to environmental parameters, it will be important to evaluate the cell response to multiple combinatorial perturbations...
January 5, 2018: Biotechnology Journal
https://www.readbyqxmd.com/read/29283217/microbioreactor-systems-for-accelerated-bioprocess-development
#17
REVIEW
Johannes Hemmerich, Stephan Noack, Wolfgang Wiechert, Marco Oldiges
In recent years, microbioreactor (MBR) systems have evolved towards versatile bioprocess engineering tools. They provide a unique solution to combine higher experimental throughput with extensive bioprocess monitoring and control, which is indispensable to develop economically and ecologically competitive bioproduction processes. MBR systems are based either on down-scaled stirred tank reactors or on advanced shaken microtiter plate cultivation devices. Importantly, MBR systems make use of optical measurements for non-invasive, online monitoring of important process variables like biomass concentration, dissolved oxygen, pH and fluorescence...
December 28, 2017: Biotechnology Journal
https://www.readbyqxmd.com/read/29283215/sequential-multivariate-cell-culture-modeling-at-multiple-scales-supports-systematic-shaping-of-a-monoclonal-antibody-towards-a-quality-target
#18
Michael Sokolov, Massimo Morbidelli, Alessandro Butté, Jonathan Souquet, Hervé Broly
The development of cell culture processes is highly complex and requires a large number of experiments on various scales to define the design space of the final process and fulfil the regulatory requirements. This work follows an almost complete process development cycle for a biosimilar monoclonal antibody, from high throughput screening and optimization to scale-up and process validation. The key goal of this analysis is to apply tailored multivariate tools to support decision-making at every stage of process development...
December 28, 2017: Biotechnology Journal
https://www.readbyqxmd.com/read/29283212/low-intensity-ultrasound-upregulates-the-expression-of-cyclin-d1-and-promotes-cellular-proliferation-in-human-mesenchymal-stem-cells
#19
Gaurav Budhiraja, Neety Sahu, Anuradha Subramanian
Human mesenchymal stem cells (hMSCs) hold great potential for cellular based therapeutics and tissue engineering applications and their expansion is an interesting prospect due to their low availability from in-vivo sources. Therefore, this study investigated the effect of continuous-wave low-intensity ultrasound (LIUS) at 5.0-MHz and 14.0-kPa (< 20 mW/cm2 ) on the proliferative capacity, colony-formation efficiency, genetic stability and differentiation potential of hMSCs. Additionally, potential signaling pathways involved in LIUS-mediated proliferation of hMSCs were studied...
December 28, 2017: Biotechnology Journal
https://www.readbyqxmd.com/read/29278307/biotransformation-of-plant-derived-phenolic-acids
#20
REVIEW
Ruchanok Tinikul, Pirom Chenprakhon, Somchart Maenpuen, Pimchai Chaiyen
Phenolic acids are abundant biomass feedstock that can be derived from the processing of lignin or other byproducts from agro-industrial waste. Although phenolic acids such as p-hydroxybenzoic acid, p-coumaric acid, caffeic acid, vanillic acid, cinnamic acid, gallic acid, syringic acid and ferulic acid can be used directly in various applications, their value can be significantly increased when they are further modified to high value-added compounds. This review summarizes and discusses the new advances in cell free and whole-cell biocatalysis technologies for reactions important for conversion of phenolic acids including esterification, decarboxylation, amination, halogenation, hydroxylation and ring-breakage reactions...
December 26, 2017: Biotechnology Journal
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