| 921. |
- Glab, Bartosz, et al.
(författare)
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Cloning of Glycerophosphocholine Acyltransferase (GPCAT) from Fungi and Plants A NOVEL ENZYME IN PHOSPHATIDYLCHOLINE SYNTHESIS
- 2016
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Ingår i: Journal of Biological Chemistry. - : AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC. - 0021-9258 .- 1083-351X. ; 291:48, s. 25066-25076
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Tidskriftsartikel (refereegranskat)abstract
- Glycero-3-phosphocholine (GPC), the product of the complete deacylation of phosphatidylcholine (PC), was long thought to not be a substrate for reacylation. However, it was recently shown that cell-free extracts from yeast and plants could acylate GPC with acyl groups from acyl-CoA. By screening enzyme activities of extracts derived from a yeast knock-out collection, we were able to identify and clone the yeast gene (GPC1) encoding the enzyme, named glycerophosphocholine acyltransferase (GPCAT). By homology search, we also identified and cloned GPCAT genes from three plant species. All enzymes utilize acyl-CoA to acylate GPC, forming lyso-PC, and they show broad acyl specificities in both yeast and plants. In addition to acyl-CoA, GPCAT efficiently utilizes LPC and lysophosphatidylethanolamine as acyl donors in the acylation of GPC. GPCAT homologues were found in the major eukaryotic organism groups but not in prokaryotes or chordates. The enzyme forms its own protein family and does not contain any of the acyl binding or lipase motifs that are present in other studied acyltransferases and transacylases. In vivo labeling studies confirm a role for Gpc1p in PC biosynthesis in yeast. It is postulated that GPCATs contribute to the maintenance of PC homeostasis and also have specific functions in acyl editing of PC (e.g. in transferring acyl groups modified at the sn-2 position of PC to the sn-1 position of this molecule in plant cells).
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| 922. |
- Glentis, G. -O, et al.
(författare)
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Efficient spectral analysis in the missing data case using sparse ML methods
- 2014
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Ingår i: European Signal Processing Conference. - 2219-5491.
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Konferensbidrag (refereegranskat)abstract
- Given their wide applicability, several sparse high-resolution spectral estimation techniques and their implementation have been examined in the recent literature. In this work, we further the topic by examining a computationally efficient implementation of the recent SMLA algorithms in the missing data case. The work is an extension of our implementation for the uniformly sampled case, and offers a notable computational gain as compared to the alternative implementations in the missing data case.
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| 923. |
- Gloriam, David E., et al.
(författare)
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A Community Standard Format for the Representation of Protein Affinity Reagents
- 2010
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Ingår i: Molecular & Cellular Proteomics. - 1535-9476 .- 1535-9484. ; 9:1, s. 1-10
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Tidskriftsartikel (refereegranskat)abstract
- Protein affinity reagents (PARs), most commonly antibodies, are essential reagents for protein characterization in basic research, biotechnology, and diagnostics as well as the fastest growing class of therapeutics. Large numbers of PARs are available commercially; however, their quality is often uncertain. In addition, currently available PARs cover only a fraction of the human proteome, and their cost is prohibitive for proteome scale applications. This situation has triggered several initiatives involving large scale generation and validation of antibodies, for example the Swedish Human Protein Atlas and the German Antibody Factory. Antibodies targeting specific subproteomes are being pursued by members of Human Proteome Organisation (plasma and liver proteome projects) and the United States National Cancer Institute (cancer-associated antigens). ProteomeBinders, a European consortium, aims to set up a resource of consistently quality-controlled protein-binding reagents for the whole human proteome. An ultimate PAR database resource would allow consumers to visit one online warehouse and find all available affinity reagents from different providers together with documentation that facilitates easy comparison of their cost and quality. However, in contrast to, for example, nucleotide databases among which data are synchronized between the major data providers, current PAR producers, quality control centers, and commercial companies all use incompatible formats, hindering data exchange. Here we propose Proteomics Standards Initiative (PSI)-PAR as a global community standard format for the representation and exchange of protein affinity reagent data. The PSI-PAR format is maintained by the Human Proteome Organisation PSI and was developed within the context of ProteomeBinders by building on a mature proteomics standard format, PSI-molecular interaction, which is a widely accepted and established community standard for molecular interaction data. Further information and documentation are available on the PSI-PAR web site. Molecular & Cellular Proteomics 9: 1-10, 2010.
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| 924. |
- Gode, Peter, et al.
(författare)
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A novel sulfonated dendritic polymer as the acidic component in proton conducting membranes
- 2006
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Ingår i: Solid State Ionics. - : Elsevier BV. - 0167-2738 .- 1872-7689. ; 177:7-8, s. 787-794
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Tidskriftsartikel (refereegranskat)abstract
- The present study involves the synthesis of sulfonated poly(3-ethyl-3-(hydroxymethyl)oxetane), sPTMPO, by end-capping the hydroxy-groups in the PTMPO with 1,4-butane sultone. A series of the polymer with different degrees of substitution was investigated. Furthermore, the subsequent use of the sulfonated PTMPO as the acidic component in proton conducting membranes was explored. The membranes were prepared by either a) mixing the partly sulfonated PTMPO with hexamethoxymethyl melamine (HMMM) to form cross-links by ether formation between the methylol groups on HMMM and the remaining hydroxyl groups on the hyperbranched polyether or b) using the sulfonated polyether in conjunction with a pyridine functionalised polysulfone, PSU-pyridine, to produce acid-base blend membranes. Membrane properties such as proton conductivity, water uptake and mechanical properties are discussed.
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| 925. |
- Gomis-Fons, Joaquín, et al.
(författare)
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Model-based design and control of a small-scale integrated continuous end-to-end mAb platform
- 2020
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Ingår i: Biotechnology progress (Print). - : John Wiley and Sons Inc.. - 8756-7938 .- 1520-6033.
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Tidskriftsartikel (refereegranskat)abstract
- A continuous integrated bioprocess available from the earliest stages of process development allows for an easier, more efficient and faster development and characterization of an integrated process as well as production of small-scale drug candidates. The process presented in this article is a proof-of-concept of a continuous end-to-end monoclonal antibody production platform at a very small scale based on a 200 ml alternating tangential flow filtration perfusion bioreactor, integrated with the purification process with a model-based design and control. The downstream process, consisting of a periodic twin-column protein A capture, a virus inactivation, a CEX column and an AEX column, was compactly implemented in a single chromatography system, with a purification time of less than 4 hr. Monoclonal antibodies were produced for 17 days in a high cell density perfusion culture of CHO cells with titers up to 1.0 mg/ml. A digital twin of the downstream process was created by modelling all the chromatography steps. These models were used for real-time decision making by the implementation of control strategies to automatize and optimize the operation of the process. A consistent glycosylation pattern of the purified product was ensured by the steady state operation of the process. Regarding the removal of impurities, at least a 4-log reduction in the HCP levels was achieved. The recovery yield was up to 60%, and a maximum productivity of 0.8 mg/ml/day of purified product was obtained.
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| 926. |
- Gomis-Fons, Joaquin, et al.
(författare)
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Optimal loading flow rate trajectory in monoclonal antibody capture chromatography
- 2021
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Ingår i: Journal of Chromatography A. - : Elsevier BV. - 0021-9673 .- 1873-3778. ; 1635
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Tidskriftsartikel (refereegranskat)abstract
- In this paper, we determined the optimal flow rate trajectory during the loading phase of a mAb capture column. For this purpose, a multi-objective function was used, consisting of productivity and resin utilization. Several general types of trajectories were considered, and the optimal Pareto points were obtained for all of them. In particular, the presented trajectories include a constant-flow loading process as a nominal approach, a stepwise trajectory, and a linear trajectory. Selected trajectories were then applied in experiments with the state-of-the-art protein A resin mAb Select PrismA (TM), running in batch mode on a standard single-column chromatography setup, and using both a purified mAb solution as well as a clarified supernatant. The results show that this simple approach, programming the volumetric flow rate according to either of the explored strategies, can improve the process economics by increasing productivity by up to 12% and resin utilization by up to 9% compared to a constant-flow process, while obtaining a yield higher than 99%. The productivity values were similar to the ones obtained in a multi column continuous process, and ranged from 0.23 to 0.35 mg/min/mL resin. Additionally, it is shown that a model calibration carried out at constant flow can be applied in the simulation and optimization of flow trajectories. The selected processes were scaled up to pilot scale and simulated to prove that even higher productivity and resin utilization can be achieved at larger scales, and therefore confirm that the trajectories are generalizable across process scales for this resin.
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| 927. |
- Gomis-Fons, Joaquin, et al.
(författare)
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Optimization study on periodic counter-current chromatography integrated in a monoclonal antibody downstream process
- 2020
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Ingår i: Journal of Chromatography A. - : Elsevier. - 0021-9673 .- 1873-3778. ; 1621
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Tidskriftsartikel (refereegranskat)abstract
- An optimization study of an integrated periodic counter-current chromatography (PCC) process in a monoclonal antibody (mAb) downstream process at lab scale, is presented in this paper. The optimization was based on a mechanistic model of the breakthrough curve in the protein-A capture step. Productivity and resin utilization were the objective functions, while yield during the loading of the capture column was set as a constraint. Different integration approaches were considered, and the effect of the feed concentration, yield and the protein-A resin was studied. The breakthrough curve and the length of the product recovery, which depended on the integration approach, determined the process scheduling. Several optimal Pareto solutions were obtained. At 0.5 mg mL−1 and 99% yield, a maximum productivity of 0.38 mg mL−1 min−1 with a resin utilization of 60% was obtained. On the other hand, the maximum resin utilization was 95% with a productivity of 0.10 mg mL−1 min−1. Due to the constraint of the process scheduling, a lower productivity could be achieved in the integration approaches with higher recovery time, which was more remarkable at higher concentrations. Therefore, it was shown that a holistic approach, where all the purification steps are considered in the process optimization, is needed to design a PCC in a downstream process.
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| 928. |
- Gorska, M., et al.
(författare)
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Evolution of the N=82 Shell Gap below 132Sn Inferred from Core Excited States in 131In
- 2009
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Ingår i: Physics Letters. Section B: Nuclear, Elementary Particle and High-Energy Physics. - : Elsevier BV. - 0370-2693 .- 1873-2445. ; 672:4-5, s. 313-316
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Tidskriftsartikel (refereegranskat)abstract
- The gamma-ray decay of ail excited state in In-131, the one proton hole neighbor of the doubly magic (132)sn has been measured. A high-spin, core-excited isomer with T-1/2 = 630(60) us was identified following production by both relativistic fragmentation of a Xe-136 beam and fission Of a U-238 beam. This state deexcites by a single gamma-ray branch of 3782(2) keV from which direct evidence for the size of the N = 82 shell gal) is inferred. The results are discussed in comparison to a shell-model Calculation including configurations across the closed shells at N = 82 and Z = 50. (C) 2009 Elsevier B.V. All rights reserved.
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| 929. |
- Gouttevin, I., et al.
(författare)
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A two-layer canopy model with thermal inertia for an improved snowpack energy balance below needleleaf forest (model SNOWPACK, version 3.2.1, revision 741)
- 2015
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Ingår i: Geoscientific Model Development. - : Copernicus GmbH. - 1991-959X .- 1991-9603. ; 8:8, s. 2379-2398
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Tidskriftsartikel (refereegranskat)abstract
- A new, two-layer canopy module with thermal inertia as part of the detailed snow model SNOWPACK (version 3.2.1) is presented and evaluated. As a by-product of these new developments, an exhaustive description of the canopy module of the SNOWPACK model is provided, thereby filling a gap in the existing literature. In its current form, the two-layer canopy module is suited for evergreen needleleaf forest, with or without snow cover. It is designed to reproduce the difference in thermal response between leafy and woody canopy elements, and their impact on the underlying snowpack or ground surface energy balance. Given the number of processes resolved, the SNOWPACK model with its enhanced canopy module constitutes a sophisticated physics-based modeling chain of the continuum going from atmosphere to soil through the canopy and snow. Comparisons of modeled sub-canopy thermal radiation to stand-scale observations at an Alpine site (Alptal, Switzerland) demonstrate improvements induced by the new canopy module. Both thermal heat mass and the two-layer canopy formulation contribute to reduce the daily amplitude of the modeled canopy temperature signal, in agreement with observations. Particularly striking is the attenuation of the nighttime drop in canopy temperature, which was a key model bias. We specifically show that a single-layered canopy model is unable to produce this limited temperature drop correctly. The impact of the new parameterizations on the modeled dynamics of the sub-canopy snowpack is analyzed. The new canopy module yields consistent results but the frequent occurrence of mixed-precipitation events at Alptal prevents a conclusive assessment of model performance against snow data. The new model is also successfully tested without specific tuning against measured tree temperature and biomass heatstorage fluxes at the boreal site of Norunda (Sweden). This provides an independent assessment of its physical consistency and stresses the robustness and transferability of the chosen parameterizations. The SNOWPACK code including the new canopy module, is available under Gnu General Public License (GPL) license and upon creation of an account at https://models.slf.ch/.
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| 930. |
- Graves, Alex, et al.
(författare)
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An experimental and theoretical investigation on Ti-5553/WC-Co(6%) chemical interactions during machining and in diffusion couples
- 2023
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Ingår i: Wear. - : Elsevier BV. - 0043-1648 .- 1873-2577. ; 516-517, s. 204604-
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Tidskriftsartikel (refereegranskat)abstract
- Chemical interactions that drive crater wear in turning are often studied using diffusion couples where the tool and workpiece are fixed. In contrast, in actual turning, there is a constant supply of new workpiece material at the tool-chip interface. In this work, diffusion simulations of a WC-Co(6%) and Ti-5Al-5V system were conducted, with constant replenishment of titanium at the interface (open system) and a fixed amount of material (closed system). The simulations showed that the formation of W(bcc), ry-phase, and TiC is dependent on the activity of C and the permeability of Co and C in titanium. Scanning and transmission electron microscopy-based techniques were used to analyse a Ti-5Al-5V-5Mo-3Cr and WC-Co(6%) diffusion couple and a worn WC-Co(6%) insert. The sequence of phases in the closed system simulation was similar to that observed in the diffusion couple. The open system simulation indicated that W(bcc) can form at WC-WC boundaries (where Co is low) within the subsurface of a WC-Co(6%) that has adhered titanium, and at the WC/Ti interface. Additionally, high densities of stacking faults and dislocations were found within subsurface WC grains, indicating a significant reduction of the tool's integrity.
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