| 1. |
- Ajjan, Fátima, et al.
(författare)
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Enhancing Energy Storage Devices with Biomacromolecules in Hybrid Electrodes
- 2019
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Ingår i: Biotechnology Journal. - : WILEY-V C H VERLAG GMBH. - 1860-6768 .- 1860-7314. ; 14:12
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Forskningsöversikt (refereegranskat)abstract
- The development of energy storage devices with higher energy and power outputs, and long cycling stability is urgently required in the pursuit of the expanding challenges of electrical energy storage. The utilization of biologically renewable redox compounds holds a great potential in designing sustainable energy storage systems and contributes in reducing the dependence on fossil fuels for energy materials. Quinones are the principal redox centers in natural organic materials and play a key role as charge storage electrode materials because of their abundance, multiple forms and integration into the materials flow through the biosphere. Electrical energy storage devices and systems can be significantly improved by the combination of scalable quinone-based biomaterials with good electronic conductors. This review uses recent examples to show how biopolymers are providing new directions in the development of renewable biohybrid electrodes for energy storage devices.
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| 2. |
- Alinaghi, Masoumeh, et al.
(författare)
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Hierarchical time-series analysis of dynamic bioprocess systems
- 2022
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Ingår i: Biotechnology Journal. - : John Wiley & Sons. - 1860-6768 .- 1860-7314. ; 17:12
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Tidskriftsartikel (refereegranskat)abstract
- Background: Monoclonal antibodies (mAbs) are leading types of ‘blockbuster’ biotherapeutics worldwide; they have been successfully used to treat various cancers and chronic inflammatory and autoimmune diseases. Biotherapeutics process development and manufacturing are complicated due to lack of understanding the factors that impact cell productivity and product quality attributes. Understanding complex interactions between cells, media, and process parameters on the molecular level is essential to bring biomanufacturing to the next level. This can be achieved by analyzing cell culture metabolic levels connected to vital process parameters like viable cell density (VCD). However, VCD and metabolic profiles are dynamic parameters and inherently correlated with time, leading to a significant correlation without actual causality. Many time-series methods deal with such issues. However, with metabolic profiling, the number of measured variables vastly exceeds the number of experiments, making most of existing methods ill-suited and hard to interpret. Methods and MajorResults: Here we propose an alternative workflow using hierarchical dimension reduction to visualize and interpret the relation between evolution of metabolic profiles and dynamic process parameters. The first step of proposed method is focused on finding predictive relation between metabolic profiles and process parameter at all time points using OPLS regression. For each time point, the p(corr) obtained from OPLS model is considered as a differential metabogram and is further assessed using principal components analysis (PCA).Conclusions: Compared to traditional batch modeling, applying proposed methodology on metabolic data from Chinese Hamster Ovary (CHO) antibody production characterized the dynamic relation between metabolic profiles and critical process parameters.
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| 3. |
- Alm, Tove, et al.
(författare)
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High-throughput protein purification under denaturating conditions by the use of cation exchange chromatography
- 2007
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Ingår i: Biotechnology Journal. - : Wiley. - 1860-6768 .- 1860-7314. ; 2, s. 709-716
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Tidskriftsartikel (refereegranskat)abstract
- A high-throughput protein purification strategy using the polycationic Z(basic) tag has been developed. In order for the strategy to be useful both for soluble and less soluble proteins, a denaturating agent, urea, was used in all purification steps. First, four target proteins were genetically fused to the purification tag, Z(basic). These protein constructs were purified by cation exchange chromatography and eluted using a salt gradient. From the data achieved, a purification strategy was planned including stepwise elution to enable parallel protein purification using a laboratory robot. A protocol that includes all steps, equilibration of the chromatography resin, load of sample, wash, and elution, all without any manual handling steps, was handled by the laboratory robot. The program allows automated purification giving milligram amounts of pure recombinant protein of up to 60 cell lysates. In this study 22 different protein constructs, with different characteristics regarding pI and solubility, were successfully purified by the laboratory robot. The data show that Z(basic) can be used as a general purification tag also under denaturating conditions. Moreover, the strategy enables purification of proteins with different pI and solubility using ion exchange chromatography (IEXC). The procedure is highly reproducible and allows for high protein yield and purity and is therefore a good complement to the commonly used His(6)-tag.
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| 4. |
- Andersson, Ken G., et al.
(författare)
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Autotransporter-Mediated Display of a Naive Affibody Library on the Outer Membrane of Escherichia coli
- 2019
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Ingår i: Biotechnology Journal. - : WILEY-V C H VERLAG GMBH. - 1860-6768 .- 1860-7314. ; 14:4
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Tidskriftsartikel (refereegranskat)abstract
- Development of new affinity proteins using combinatorial protein engineering is today established for generation of monoclonal antibodies and also essential for discovery of binders that are based on non-immunoglobulin proteins. Phage display is most frequently used, but yeast display is becoming increasingly popular, partly due to the option of utilizing fluorescence-activated cell sorting (FACS) for isolation of new candidates. Escherichia coli has several valuable properties for library applications and in particular the high transformation efficiency. The use of various autotransporters and intimins for secretion and anchoring on the outer membrane have shown promising results and particularly for directed evolution of different enzymes. Here, the authors report on display of a large naive affibody library on the outer membrane of E. coli using the autotransporter Adhesin Involved in Diffuse Adherence (AIDA-I). The expression cassette is first engineered by removing non-essential sequences, followed by introduction of an affibody library, comprising more than 10(9) variants, into the new display vector. The quality of the library and general performance of the method is assessed by FACS against five different targets, which resulted in a panel of binders with down to nanomolar affinities, suggesting that the method has potential as a complement to phage display for generation of affibody molecules.
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| 5. |
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| 6. |
- Borodina, I., et al.
(författare)
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Advances in metabolic engineering of yeast Saccharomyces cerevisiae for production of chemicals
- 2014
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Ingår i: Biotechnology journal. - : Wiley. - 1860-6768 .- 1860-7314. ; 9:5, s. 609-620
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Forskningsöversikt (refereegranskat)abstract
- Yeast Saccharomyces cerevisiae is an important industrial host for production of enzymes, pharmaceutical and nutraceutical ingredients and recently also commodity chemicals and biofuels. Here, we review the advances in modeling and synthetic biology tools and how these tools can speed up the development of yeast cell factories. We also present an overview of metabolic engineering strategies for developing yeast strains for production of polymer monomers: lactic, succinic, and cis,cis-muconic acids. S. cerevisiae has already firmly established itself as a cell factory in industrial biotechnology and the advances in yeast strain engineering will stimulate development of novel yeast-based processes for chemicals production.
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| 7. |
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| 8. |
- Braissant, Olivier, et al.
(författare)
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Isothermal microcalorimetry accurately detects bacteria, tumorous microtissues, and parasitic worms in a label-free well-plate assay
- 2015
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Ingår i: Biotechnology Journal. - : Wiley. - 1860-6768 .- 1860-7314. ; 10:3, s. 460-468
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Tidskriftsartikel (refereegranskat)abstract
- Isothermal microcalorimetry is a label-free assay that allows monitoring of enzymatic and metabolic activities. The technique has strengths, but most instruments have a low throughput, which has limited their use for bioassays. Here, an isothermal microcalorimeter, equipped with a vessel holder similar to a 48-well plate, was used. The increased throughput of this microcalorimeter makes it valuable for biomedical and pharmaceutical applications. Our results show that the sensitivity of the instrument allows the detection of 3 x 10(4) bacteria per vial. Growth of P. mirabilis in Luria Broth medium was detected between 2 and 9 h with decreasing inoculum. The culture released 2.1J with a maximum thermal power of 76 W. The growth rate calculated using calorimetric and spectrophotometric data were 0.60 and 0.57 h(-1), respectively. Additional insight on protease activities of P. mirabilis matching the last peak in heat production could be gathered as well. Growth of tumor microtissues releasing a maximum thermal power of 2.1 W was also monitored and corresponds to a diameter increase of the microtissues from ca. 100 to 428 m. This opens new research avenues in cancer research, diagnostics, and development of new antitumor drugs. For parasitic worms, the technique allows assessment of parasite survival using motor and metabolic activities even with a single worm.
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| 9. |
- Campbell, Kate, 1987, et al.
(författare)
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Cell-to-cell heterogeneity emerges as consequence of metabolic cooperation in a synthetic yeast community
- 2016
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Ingår i: Biotechnology journal. - : Wiley. - 1860-6768 .- 1860-7314. ; 11:9, s. 1169-1178
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Tidskriftsartikel (refereegranskat)abstract
- Cells that grow together respond heterogeneously to stress even when they are genetically similar. Metabolism, a key determinant of cellular stress tolerance, may be one source of this phenotypic heterogeneity, however, this relationship is largely unclear. We used self-establishing metabolically cooperating (SeMeCo) yeast communities, in which metabolic cooperation can be followed on the basis of genotype, as a model to dissect the role of metabolic cooperation in single-cell heterogeneity. Cells within SeMeCo communities showed to be highly heterogeneous in their stress tolerance, while the survival of each cell under heat or oxidative stress, was strongly determined by its metabolic specialization. This heterogeneity emerged for all metabolite exchange interactions studied (histidine, leucine, uracil, and methionine) as well as oxidant (H2O2, diamide) and heat stress treatments. In contrast, the SeMeCo community collectively showed to be similarly tolerant to stress as wild-type populations. Moreover, stress heterogeneity did not establish as sole consequence of metabolic genotype (auxotrophic background) of the single cell, but was observed only for cells that cooperated according to their metabolic capacity. We therefore conclude that phenotypic heterogeneity and cell to cell differences in stress tolerance are emergent properties when cells cooperate in metabolism.
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| 10. |
- Camsund, Daniel, et al.
(författare)
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Genetically engineered light sensors for control of bacterial gene expression
- 2011
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Ingår i: Biotechnology Journal. - : Wiley. - 1860-6768 .- 1860-7314. ; 6:7, s. 826-836
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Forskningsöversikt (refereegranskat)abstract
- Light of different wavelengths can serve as a transient, noninvasive means of regulating gene expression for biotechnological purposes. Implementation of advanced gene regulatory circuits will require orthogonal transcriptional systems that can be simultaneously controlled and that can produce several different control states. Fully genetically encoded light sensors take advantage of the favorable characteristics of light, do not need the supplementation of any chemical inducers or co-factors, and have been demonstrated to control gene expression in Escherichia coli. Herein, we review engineered light-sensor systems with potential for in vivo regulation of gene expression in bacteria, and highlight different means of extending the range of available light input and transcriptional output signals. Furthermore, we discuss advances in multiplexing different light sensors for achieving multichromatic control of gene expression and indicate developments that could facilitate the construction of efficient systems for light-regulated, multistate control of gene expression.
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